J. Biosci., Vol. 22, Number 3, June 1997, pp 367–374. © Printed in India. Karyology of a few species of south Indian acridids. II Male germ line karyotypic instability in Gastrimargus§ H CHANNAVEERAPPA* and H A RANGANATH† Department of Studies in Zoology, University of Mysore, Manasagangotri, Mysore 570006, India *Government College for Women, Mandya 571401, India MS received 26 April 1996; revised 20 September 1996 Abstract. Gastrimargus africanus orientalis, an acridid grasshopper has revealed the existence of karyotypic mosaicism in the male germ line cells of a few individuals with 2n = 23, 19, 21, 25 and 27 chromosomes. Details of this chromosomal instability are presented in this paper. Keywords. Karyology; south Indian acridids; male germ line; karyotypic instability; Gastrimargus africanus orientalis. 1. Introduction The karyotype is an important asset of a species. In spite of 'polymorphism', the architecture and composition of a species karyotype is relatively stable. The karyology of acridid grasshoppers presents a very stable phenotype over a wide range of species. It is often quoted as an example for "karyotypic conservatism" (White 1973; Ashwath 1981; Yadav and Yadav 1993). But Gastrimargus, an acridid, illustrates a facet of 'karyotypic dynamism' with a rare phenomenon of aneuploids in the male germ line and the same is presented in this paper. 2. Materials and methods The individuals of Gastrimargus africanus orientalis Saussure were collected from Mandya (Karnataka) during 1993-95. A total of 128 individuals (94 males and 34 females) were analised. Adult testis/ovarian follicles and intestinal caecae were removed and fixed in 3:1 methanol and glacial acetic acid. Squash preparations were made without colchicine and hypotonic treatments, and the material was stained with Heidenhain's iron haematoxylin stain. Well spread mitotic and meiotic stages were analysed and photographed. 3. Results and discussion The karyology of every species is unique to itself and provides an identity to the species (Channaveerappa and Ranganath 1994). The study of karyotypic evolution in different §We dedicate this paper to our teacher Prof. L Siddaveere Gowda on the eve of his 60th birthday. †Corresponding author. 367 368 Η Channaveerappa and Η A Ranganath groups of animals, has revealed that the karyotype is not absolutely stable, instead changes do occur on an evolutionary time scale. In this regard, Gastrimargus has attracted the attention of cytologists eversince 1963. Rajasekarasetty (1963) reported that in addition to normal complement of 2n = 23, the metaphase I of a few cells had 9 bivalents and an X-chromosome, indicating the diploid number as 2n = 19. But the later studies of Ashwath (1981), Ashwathanarayana et al (1981) in G. a. orientalis; Fossey and Liebenberg (1991) in four species of Gastrimargus, namely G. africanus, G. crassicollis, G. vitripennisandG. wahlbergi;Johnetal(1985) inG. musicusand Yadav and Yadav (1993) in G. transversus have not revealed a situation similar to the one, reported by Rajasekarasetty (1963). The present analysis of G. a. orientalis has revealed that both the somatic and the germ line cells of all females (N=34) analysed had a karyotype of 2n = 24 (22AA + XX). On the other hand, 82 of the 94 males analysed had 2n = 23 (22AA + X) in both somatic and germ line cells (figure 1). This is typical of acridid grasshoppers with XX female and XO male systems. The autosome complement includes 3 pairs of long, 6 pairs of medium and 2 pairs of short chromosomes. The X-chromosome is Figure 1. (A) Gonial metaphase of male with normal complement of 2n = 23 (22AA + X). (B) Karyotype of normal male complement (2n = 23). Karyotypic instability of male germ line in Gastrimargus 369 slightly larger th an the largest autosomal pair. This is similar to the earlier reports on the karyotype of G. a. orientalis (Ashwathanarayana et al 1981). Further, the present study has revealed that except the 6th, 9th, 10th and 1lth pair of autosomes, rest of the autosomes and the sex chromosomes are acrocentric, while these pairs are polymer- phic with acrocentric, metacentric and submetacentric representatives, which will be reported elsewhere. The chromosomal constitution of the other 12 males of Gastrimargus under study is interesting. The somatic cells (intestinal caecae had 2n = 23. The germinal tissue of four males had four types of cells, viz., 2n = 23, 19, 21 and 25 (figures 2 and 3), while the other 8 males had five types of cells. In addition to the above four types, also had cells with 2n = 27 (figures 2 and 3). Thus the germinal tissue of these 12 males of Gastrimargus Figure 2. Metaphase plates with different diploid number of chromosomes. (A) 2n = 19. (B) 2n = 21.(C) 2n = 25. (D) 2n = 27. 370 Η Channaveerappa and Η A Ranganath Figure 3. Karyotypes of different diploid complements. (A) 2n = 19 (8th and 10th pairs absent). (B) 2n = 21 (8th pair absent). (C) 2n = 25 (10th pair duplicated). (D) 2n = 27 (8th and 9th pairs duplicated). Karyotypic instability of male germ line in Gastrimargus 371 Table 1. The incidence of cells with different numbers of chromosomes in 12 males of G. a. orientalis. The values within brackets are the percentages. Note: Variant karyotypes are observed in all the follicles of testis in each individual. show 'karyotypic mosaicism' (table 1). The cells with normal complement of 2n = 23 preponderate over other category. In individuals which had up to 2n = 27, cells with different aneuploid composition have occurred in almost equal proportions while in individuals which had 2n = 25, cells with 2n = 19 and 2n = 21 are more frequent than cells with 2n = 25. The standard karyotype with 2n = 23 carries every pair of chromosome only once as expected (figure 1). But in the aneuploid cells of G. a. orientalis it varies. For instance, 2n = 19, is due to the absence of 8th and 10th pairs (figures 2A, 3A); 2n = 21 is due to the absence of 8th or 9th pair of chromosomes (figures 2B, 3B). In 2n = 25, the 8th or the 10th pairs were represented twice (figures 2C, 3C) while 2n = 27 has resulted due to the duplication of 8th and 9th pairs or 9th and 10th pairs of chromosomes (figures 2D, 3D). Therefore, it appears that the numerical variation centres around 8th, 9th and 10th pairs of chromosomes. Such numerical variations are also noticed in subsequent meiotic stages (figures 4A-4D, 5A-5C). Thus far there is no indication of the involve- ment of the X-chromosome in this phenomena in G. orientalis. A deviation from the standard chromosome complement within an individual is confined to germ line cells of a few males only, while its somatic cells (gastric caecae) were found to carry only the Standard complement. So far such an intraindivi- dual variation is not seen in females of G. a. orientalis. The preliminary report of Rajasekarasetty (1963) had revealed only one type of aneuploid (2n = 19) that too in metaphase I and anaphase of G. a. orientalis. In contrast, the present study had revealed a wide range of aneuploid through various stages of meiosis. Significant variations in chromosome number involving Standard member of the karyotype is rare. Lewis and John (1959) in a locust, and Sharma et al (1965) in Chrotogonus trachypterus, had shown that polysomy in male germ line is due to different members of the complement. But in Chorthippus parallelus (Hewitt and John 1968) the polysomy was found to be due to the same chromosome. Similarly in Atractomorpha similis (Peters 1981), a few males were found to carry between 1 to 10 extra copies of a particular chromosome. Similar situations were also seen in Gomphocerus Sibiriens (Gosalvez and Lopez-Fernandez 1981), Omocestus bolivari (Viseras and Camacho 1982), and Chorthippus binotatus (Talavera et al 1990). These earlier findings revealed the occurrence of 'additional copies' of the chromosome of a normal complement. On the contrary, in the present investigations in G. a. orientalis, deviation was due to not only to extra representation but also was due to lack of total 372 H Channaveerappa H A Ranganath Figure 4. Meiotic stages in aneuploids. Diakinesis with (A) 9, (B) 10, (C) 12 and (D) 13 bivalents + X. representation of a pair(s) of chromosomes. In G. a. orientalis, thus far there is no indication of one of the members of a pair absent or represented in more copies. Therefore monosomic or trisomic cells etc., were not found. This important observa- tion indicates that presumably only specific types of chromosome malsegregation of 8th, 9th and 10th pairs in specific germ cells do occur. With the present data, it is difficult to visualize the possible mechanism(s) that underline this karyotypic Karyotypic instability of male germ line in Gastrimargus 373 Figure 5. Meiotic stages in aneuploids. Metaphase II with (A) 10 chromosomes (9A + X) and (B) 9 chromosomes (9A). (C) Anaphase I with 11 and 10 chromosomes in two poles. instability. Recently, Ford and Correll (1992) have studied the errors in mitotic divisions by using satellite DNA as probes and found to be due to non-disjunction, chromosome and chromatid lagging, chromatid mal segregation and monopolar segre- gation. The significance of this karyotypic mosaicism is difficult to hypothesize. That this rare type of instability exclusively seen in a few cells of a few individuals of a population may be an evolutionary experimentation with male germline which can withstand the impact of such aneuplody. Further studies are in progress to understand the influence of karyotypic mosaicism in G. a. orientalis on spermatids, sperm morphology, fertility, etc. 374 Η Channaveerappa and Η A Ranganath Acknowledgements Authors are grateful to the Chairman, Department of Studies in Zoology, Manasagangotri, Mysore for facilities. HC is indebted to the Director, Collegiate Education, Government of Karnataka and to the Principal, Government College for Women, Mandya for permission and to Dr Η Ρ Puttaraju for suggestions.